Possible role of autoimmunity in patients with premature ovarian insufficiency.

BACKGROUND: To evaluate the involvement of immune abnormality in patients with idiopathic premature ovarian insufficiency (POI). In addition to the known etiology, autoimmune disorders may be a pathologic mechanism for POI. MATERIALS AND METHODS: Our study was a prospective controlled trial. Twenty women with POI, reasons other than autoimmune excluded, were enrolled in this study. The control group consisted of 17 healthy women. In both groups, family and personal history were taken and the levels of follicle stimulating hormone, luteinizing hormone, thyroid-stimulating hormone, prolactin, anti-Müllerian hormone, inhibin B, antithyroglobulin and antithyroid peroxidase antibodies were determined. Antiovarian antibodies and subpopulations of peripheral blood T-lymhocytes were also determined. RESULTS: Participants in the study group exhibited hypergonadotropichypogonadism, while high levels of follicle stimulating hormone and low levels of inhibin B and anti-Müllerian hormone were observed. In 16 (80%) patients, POI was associated in their personal and familial history with another autoimmune disease. Fifty percent of patients presented highly elevated antithyroid antibodies. The lymphocyte subset, especially B cells, was significantly higher (p=0.014), and peripheral regulatory lymphocytes CD25+ high were significantly lower (p=0.015) in the study group than in the control group. Anti- ovarian antibodies were detected in 20% of patients with POI. CONCLUSION: We presume that the presence of anti-ovarian antibodies together with abnormalities of cellular immunity may in some cases potentially represent the involvement of an autoimmune mechanism in idiopathic POI.

Expression of pluripotency and oocyte-related genes in single putative stem cells from human adult ovarian surface epithelium cultured in vitro in the presence of follicular fluid.

The aim of this study was to trigger the expression of genes related to oocytes in putative ovarian stem cells scraped from the ovarian surface epithelium of women with premature ovarian failure and cultured in vitro in the presence of follicular fluid, rich in substances for oocyte growth and maturation. Ovarian surface epithelium was scraped and cell cultures were set up by scrapings in five women with nonfunctional ovaries and with no naturally present mature follicles or oocytes. In the presence of donated follicular fluid putative stem cells grew and developed into primitive oocyte-like cells. A detailed single-cell gene expression profiling was performed to elucidate their genetic status in comparison to human embryonic stem cells, oocytes, and somatic fibroblasts. The ovarian cell cultures depleted/converted reproductive hormones from the culture medium. Estradiol alone or together with other substances may be involved in development of these primitive oocyte-like cells. The majority of primitive oocyte-like cells was mononuclear and expressed several genes related to pluripotency and oocytes, including genes related to meiosis, although they did not express some important oocyte-specific genes. Our work reveals the presence of putative stem cells in the ovarian surface epithelium of women with premature ovarian failure.

Effect of counseling on adherence to perimenopausal hormone therapy in Slovenia.

OBJECTIVE: To evaluate the effect of counseling on women's adherence to hormone therapy (HT) in Slovenia following the publication of the results of the Women's Health Initiative prospective study. METHOD: In this 24-month prospective, randomized, controlled study with 125 women in early menopause, those in the study group (n=64) attended oral presentations on menopause and HT whereas those in the control group (n=63) did not. Data were collected from 2 questionnaires, one completed before starting HT and the other during follow-up visits after 3, 6, 12, and 24 months. The results were analyzed according to the intention-to-treat principle. RESULTS: In the second year, an improved quality of life and friends critical of HT negatively affected adherence to HT. After 24 months, 47% of the participants in the study group and 32% in the control group were still using HT. The difference between the 2 groups in the rates of participants who discontinued HT was the same in the first than and in the last 12 months of the study (49 [76.6%] vs 15 [23.4%]). CONCLUSION: Since there were no changes in discontinuation rates between the groups from one study period to the other, and a total of 15 women in each group stopped using HT in the second year of the study, the effect of the educational presentations on HT adherence was long-term rather than temporary.

An observational study of assisted reproductive technology outcomes in new European Union member states: an overview of protocols used for ovarian stimulation.

BACKGROUND: The development of new fertility treatment options has facilitated individualized assisted reproductive technology (ART) protocols to improve outcomes. Manufacturing improvements to recombinant human follitropin alfa have allowed precise dosing based on mass (filled-by-mass; FbM) rather than bioactivity (filled-by-bioassay; FbIU). Continued monitoring and reporting of follitropin alfa treatment outcomes in routine clinical practice is essential. OBJECTIVE: To provide an overview of the frequency of different controlled ovarian-stimulation protocols used in in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles in new European Union member states, and to provide post-registration efficacy and safety data on follitropin alfa. RESEARCH DESIGN AND METHODS: A 2-year, prospective, observational, multicentre, Phase IV study conducted at ART clinics in the Czech Republic, Estonia, Latvia, Lithuania, Poland, Slovakia and Slovenia. Women aged 18-47 years undergoing ovarian stimulation with follitropin alfa for conventional IVF or ICSI were eligible for inclusion. The main treatment outcome was cumulative clinical pregnancy rate. Data were analysed descriptively. RESULTS: Clinical pregnancy outcomes were available for 4055 of 4085 (99.3%) patients. In total, 1897 (46.8%) patients used follitropin alfa FbIU; 2133 (52.6%) used follitropin alfa FbM. Clinical pregnancy was achieved by 39.5% (1603/4055) of patients. A greater proportion of patients with polycystic ovary syndrome achieved a clinical pregnancy than those with endometriosis (41.8% vs 37.8%, respectively). A higher cumulative pregnancy rate was observed with the use of follitropin alfa FbM than follitropin alfa FbIU (41.3% vs 37.8%, respectively; p = 0.02). CONCLUSIONS: This study represents the most comprehensive audit of individualized ART in clinical practice in Central and Eastern Europe. Overall, clinical pregnancy was achieved by 39.5% of patients after stimulation with follitropin alfa. The use of follitropin alfa FbM resulted in a higher cumulative pregnancy rate than did the FbIU formulation. However, limitations of the study include the observational and non-comparative study design, and descriptive nature of statistical analyses; furthermore, the study was not designed to make direct comparisons between the success rates of different ovarian-stimulation protocols.

Male and female factors that influence ICSI outcome in azoospermia or aspermia.

Factors that influenced the clinical results of 220 first-attempt intracytoplasmic sperm injection (ICSI) cycles with testicular spermatozoa were evaluated in 107 men with non-obstructive azoospermia, 72 with obstructive azoospermia and 41 with aspermia. Linear and logistic regression analysis showed that the fertilization rate depended positively on Johnsen score (P = 0.016) and on the type of ovarian stimulation: a higher fertilization rate was observed after ovarian stimulation with agonist and recombinant FSH than after stimulation with agonist and urinary menopausal gonadotrophin (P = 0.026). Embryo development to the blastocyst stage was predicted positively by the number of injected oocytes (P = 0.016) and negatively by male FSH concentration (P = 0.019). A higher proportion of blastocysts developed after the use of frozen-thawed spermatozoa in comparison to fresh spermatozoa (P = 0.034). Embryo development to the blastocyst stage influenced pregnancy (P = 0.002) and live birth outcomes (P = 0.005); live birth was also predicted by female age (P = 0.046). Embryo culture to day 5 in comparison to day 2 did not provide higher live birth rates. In azoospermia/aspermia, the ICSI outcome depends on both male factors (FSH, Johnsen score, sperm status and motility) and female factors (age, number of injected oocytes).

Parthenogenetic embryo-like structures in the human ovarian surface epithelium cell culture in postmenopausal women with no naturally present follicles and oocytes.

Little is known about parthenogenesis in the human ovary. What is known is related to patients with teratoma in their medical history. Ovarian surface epithelium (OSE) was often proposed as a source of ovarian stem cells with an embryonic character in the past, and was also termed "germinal epithelium." The aim of this study was to isolate putative stem cells from OSE scrapings, to set up an OSE cell culture, to follow the in vitro oogenesis and possible formation of parthenogenetic embryos in 21 postmenopausal women with no naturally present follicles and oocytes. Small round cells with a bubble-like structure and with a diameter from 2 to 4 microm were isolated from the material obtained by OSE scrapings in all women. They expressed early embryonic developmental markers such as stage-specific embryonic antigen-4 (SSEA-4) surface antigen and Oct-4, Nanog, Sox-2, and c-kit transcription factors. These cells were separated by density gradient centrifugation and grown in vitro, where they proliferated and formed embryoid body-like structures. Their markers of pluripotency such as telomerase activity were decreased during in vitro culture and they did not form teratoma after the injection into SCID mice. Some of them grew intensively and reached a diameter of approximately 20 microm after 5-7 days of culture. In the OSE cell culture, oocyte-like cells developed among them, which reached a diameter up to 95 mum, and expressed Oct-4, c-kit, VASA, and ZP2 transcription markers after 20 days of culture. Some of them expressed a zona pellucida-like structure and rarely germinal vesicle- and polar body-like structures. At the same time, parthenogenetic blastocyst-like structures developed, which expressed transcription markers Oct-4, Sox-2, and Nanog and were normal for chromosomes X, Y, 13, 16, 18, 21, and 22. In conclusion, the discovered cells expressed embryonic stem cell markers, gave rise to embryoid body-, oocyte-, and blastocyst-like structures, and might be involved in the human reproduction and formation of ovarian tumors.

Putative stem cells with an embryonic character isolated from the ovarian surface epithelium of women with no naturally present follicles and oocytes.

There have been some proposals that stem cells exist in the ovarian surface epithelium (OSE) of the adult human ovary; however, no direct evidence of such cells has been given until now. The aim of this study was to isolate the putative ovarian stem cells (OSCs) from the OSE layer in women with no naturally present oocytes and follicles--20 postmenopausal women and five women with premature ovarian failure. Small round cells with a bubble-like structure and diameters from 2 to 4 microm were isolated from the material obtained by OSE scraping. They expressed early embryonic developmental markers such as stage-specific embryonic antigen-4 and Oct-4, Nanog, Sox-2, and c-kit transcription markers, and they displayed prominent c-kit immunohistochemical staining. These cells were separated by density gradient centrifugation and grown in vitro, where they proliferated. Some of them grew intensively and reached a diameter of approximately 20 microm after 5-7 days. In the OSE cell culture, oocyte-like cells developed, which reached a diameter of up to 95 microm and expressed Oct-4A, Oct-4B, c-kit, VASA, and ZP2 transcription markers, corresponding to early oocytes. They did not express SCP3 meiotic marker. In conclusion, the discovered cells are proposed to represent the adult OSCs with the expression of embryonic stem cell markers. The expression of germ lineage marker c-kit points toward their primordial germ cell ancestry. A new term "embryonic-like stem cells of the adult" is proposed for embryonic-like stem cells that might persist in various tissues and organs of adults. These findings could be used for further studies aimed at the autologous treatment of ovarian infertility and degenerative diseases.

Single fresh and frozen-thawed blastocyst transfer using hyaluronan-rich transfer medium.

The purpose of this prospective randomized study was to evaluate use of a hyaluronan-rich transfer medium in fresh and frozen-thawed single blastocyst transfer. The study included 279 single blastocyst transfers in women aged<37 years in their first, second or third treatment cycle. According to the type of single blastocyst transfer (fresh elective or frozen-thawed) the women were divided into two study and two control groups. In both study groups (n=130) transfers were performed using hyaluronan and in the control groups (n=149) a conventional transfer medium was used. The results indicate that fresh elective single blastocyst transfer with hyaluronan results in significantly higher pregnancy rates in a selected subgroup of women; those with >or=2 blastocysts developed to day 5 and a previous implantation failure (55% versus 10%; P=0.012). Overall pregnancy rates after fresh elective and frozen-thawed single blastocyst transfer were similar in both study and control groups.

Decreased androgen levels and improved menstrual pattern after angiotensin II receptor antagonist telmisartan treatment in four hypertensive patients with polycystic ovary syndrome: case series.

We describe 4 consecutive hypertensive women with polycystic ovary syndrome, classified according to the National Institute of Child Health and Human Development (NICHD) criteria, treated with telmisartan 40 mg/d for six months. Blood pressure, menstrual pattern, body mass index (BMI), homeostasis model assessment of insulin resistance, testosterone, dehydroepiandrosterone sulfate (DHEAS), and androstenedione were recorded and measured before and after telmisartan treatment. Obese hypertensive polycystic ovary syndrome patients had a decrease in systolic blood pressure. Marked drop-off in serum androgen concentrations was observed in all four patients. Three patients improved their menstrual cyclicity. The improvements were independent of changes in weight. The reduction of androgen concentrations and improvement in menstrual pattern was achieved despite a non-significant change of fasting insulin levels in patients, who were not considered severely insulin resistant at baseline. These findings may provide a new basis for a proper choice of the antihypertensive drug in hypertensive women with polycystic ovary syndrome.

Polymorphisms in the interleukin-12/18 genes and recurrent spontaneous abortion.

PROBLEM: Interleukin (IL) IL-12/IL-18 are involved in uterine NK cells control of uterine vascular development. Polymorphisms in the IL-12/IL-18 genes could modify the cytokine balance, which might result in an increased susceptibility to recurrent spontaneous abortion (RSA). METHOD OF STUDY: A case-control study was conducted to determine the association between the IL12 (I/D) and IL18 (-607C>A, -137G>C) gene polymorphisms and the risk of RSA in 125 women with RSA and in 136 controls. RESULTS: The frequencies of DD, ID, II for IL-12 were, 25.6%, 52.8% and 21.6% respectively, in patients versus 21.3%, 51.5% and 27.2% respectively in controls; the frequencies of CC, CA, AA genotypes for IL-18 (-607) were, 34.4%, 54.4% and 11.2% respectively in patients versus 30.1%, 58.1% and 11.8% respectively in controls; the frequencies of GG, GC, CC genotypes for IL-18(-137) were 47.2%, 43.2% and 9.6% respectively in patients and 45.6%, 46.3% and 8.1% respectively in controls. CONCLUSION: IL-12B and IL-18 promoter gene polymorphisms were not associated with RSA in our women.

Determination of serum lysophosphatidic acid as a potential biomarker for ovarian cancer.

A fast and selective analytical method, used to determine the different lysophosphatidic acid (LPA) species in serum, has been developed and validated. LPA species were quantitatively extracted from serum using methanol-chloroform (2:1, v/v). The proteins were precipitated by this solvent mixture and separated by centrifugation in one step. LPA levels were determined in clear extracts using the HPLC-MS/MS method. The linearity of this method was established in the concentration range between 0.1 and 16 microM for all LPA species with a correlation coefficient greater than 0.99. Recovery of all LPA species determined by the serum, fortified at approximately 1 microM and 2-3 microM, was between 93% and 111% with an average R.S.D. of less than 8%. This method was used to determine LPA in numerous sera of healthy controls, patients with benign ovarian tumours and ovarian cancer at different stages. Significantly higher total LPA levels were determined in the sera of patients with different types of tumours (benign and malignant).

Leptin levels in infertile male patients are correlated with inhibin B, testosterone and SHBG but not with sperm characteristics.

In the present study, differences in leptin levels between different groups of male patients presenting with infertility problems and possible correlations between leptin levels and clinical, spermiological, histological and hormonal characteristics were examined. Two hundred and ten male partners from infertile couples were included in the study. Based on clinical examination, spermiogram and testicular histology results, patients were divided into four groups: 42 men with non-obstructive azoospermia, 15 men with obstructive azoospermia, 68 men with oligoasthenoteratozoospermia and 85 men with normozoospermia. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B, testosterone, sex hormone binding globulin (SHBG) and leptin were measured. After adjustment for body mass index, there was a negative correlation between serum levels of leptin and inhibin B, total testosterone and SHBG (r = -0.189, p = 0.009, r = -0.250, p = 0.001 and r =-0.221, p = 0.003 respectively) but there was no correlation between leptin and classical sperm characteristics. Our results therefore demonstrate a link between leptin and testicular function, independently of FSH and LH, possibly involving testosterone and SHBG through a regulation of Leydig cell function.

Outcome of frozen-thawed blastocysts derived from gonadotropin releasing hormone agonist or antagonist cycles.

PURPOSE: The aim of this retrospective study was to compare the outcome of frozen-thawed blastocysts derived from the cycles using controlled ovarian stimulation with GnRH agonists vs. GnRH antagonists. METHODS: Survival, pregnancy and cumulative live birth rates in 231 freeze-thaw cycles derived from the GnRH agonist cycles (GnRH agonist group), and in 175 freeze-thaw cycles derived from the GnRH antagonist cycles (GnRH antagonist group) were compared. RESULTS: In the GnRH agonist group significantly higher proportion of blastocysts survived the thawing procedure than in the GnRH antagonist group (86.1% versus 78.5%; p < 0.01). The differences in cumulative live birth rates did not differ significantly between the groups: in the GnRH agonist group the cumulative live birth rate was 16.5%, and in the GnRH antagonist group it was 14.2%. CONCLUSIONS: Frozen-thawed blastocysts derived from the GnRH agonist cycles have better survival rates and similar cumulative live birth rates than those derived from the GnRH antagonist cycles.

Increased ammonium in culture medium reduces the development of human embryos to the blastocyst stage.

The objective of this prospective study was to evaluate the effect of ammonium accumulated in sequential media and determined by enzymatic spectrophotometric method on the blastocyst development in 281 human embryos from 100 stimulated and natural in vitro fertilization (IVF) cycles. Ammonium concentration was increased in 62% of cycles and was correlated negatively with the blastocyst development after classical IVF, but not after intracytoplasmic sperm injection (ICSI).

Uterine arterial blood flow and the substances of ovarian renin-angiotensin system in women with polycystic ovaries undergoing in vitro fertilization.

OBJECTIVE: To find whether plasma and follicular prorenin concentrations have any effect on the uterine arterial blood flow in women with polycystic ovarian syndrome (PCOS) compared to those with normal menstrual cycles (NMC). STUDY DESIGN: Controlled prospective clinical study involved 55 women undergoing in vitro fertilization: 24 with PCOS and 31 with NMC. In both groups transvaginal colour Doppler assessment of uterine arterial blood flow was analysed on day 22 of the cycle, on the day of human chorionic gonadotrophin (HCG) administration and 36 h after HCG. Plasma and follicular (in the dominant follicle containing mature oocyte, and in the pooled follicles) prorenin and active renin, and serum estradiol and androstenedione concentrations were measured at these time-points. The Student's t-test and Pearson correlation were used for the statistical analysis. RESULTS: The resistance index (RI) in the NMC group decreased from 0.84+/-0.05 on the day of HCG to 0.78+/-0.08 36 h after HCG (P < 0.05); in the PCOS group the RI did not decrease. Follicular prorenin concentrations in the dominant follicle and in the pooled follicles were lower in the PCOS than in the NMC group (20,210+/-10,831 microU/l, 16,753+/-8634 microU/l versus 42,637+/-35,400 microU/l, 33,067+/-26,200 microU/l; P < 0.05). CONCLUSIONS: Plasma prorenin concentrations do not affect vascular impedance to the uterine artery, but follicular prorenin do by newly formed low resistant vessels in the follicles.

Continuation of hormone replacement therapy in Slovenia: a prospective, randomized, controlled trial--1-year follow-up.

OBJECTIVES: To evaluate the influence of educational intervention on the hormone replacement therapy (HRT) continuation rate in Slovenia after the publication of WHI results. METHODS: We enrolled 125 early postmenopausal women in a 12-month prospective, randomized, controlled, multicentric study in Slovenia. The study group women (n=62) attended educational lectures; the control group women (n=63) did not. Data were collected from three types of questionnaire: before starting HRT, at follow-up visits at 3, 6, 9 and 12 month, after the educational lecture (study group). The continuation rate was measured on the basis of women's self-reports. The results were analyzed according to the "intention-to-treat" principle. The Cox proportional hazard model was used for the final analysis. RESULTS: A gynecologist's suggestion, climacteric symptoms and quality of life were the prevailing reasons for starting HRT. The prevailing factors affecting continuation of HRT were: no or irregular previous OC use (hazard ratio 3.7), no educational lectures (hazard ratio 2.0) and climacteric disorders as the reason for start HRT (hazard ratio 2.1). In the women who discontinued HRT within the first 3 months, the fear of endometrial cancer, breast cancer and bleeding problems were statistically more significant than other factors (P=0.034). In the women who stopped HRT use within 6-12 months, the fear of breast and endometrial cancers increased substantially (P=0.002). CONCLUSIONS: Previous OC use and educational lectures on menopausal problems and HRT significantly improve the HRT continuation rate. The main reason for discontinuing HRT is fear of breast cancer, intensified by media.

Intracytoplasmic sperm injection as a complement to gonadotrophin treatment in infertile men with hypogonadotrophic hypogonadism.

In this study we sought to determine whether intracytoplasmic sperm injection (ICSI) could improve the efficacy of treatment with gonadotrophins in gonadotrophin-deficient men in terms of pregnancy. A series of six adult men (aged 26-47 years) with hypogonadotrophic hypogonadism (HH) is reported: four men with prepubertal isolated idiopathic HH (IIHH) and two adult-onset HH, as part of hypopituitarism secondary to surgical treatment of a pituitary tumour. All were azoospermic. To restore spermatogenesis, all received hormonal treatment with intramuscular human menopausal gonadotrophins (HMG) and human chorionic gonadotrophin (HCG) for 2 to 23 months. High basal serum inhibin B was predictive of rapid and complete recovery of spermatogenesis. In the two adult-onset HH, a natural pregnancy was achieved within 3 months. The four men with IIHH underwent ICSI because of poor sperm quality. ICSI using fresh or frozen-thawed ejaculated spermatozoa was performed after 6-23 months of gonadotrophin treatment. ICSI provided good clinical results in terms of fertilization and embryo quality, and resulted in three pregnancies that ended in three term deliveries. In men with oligozoospermia related to prepubertal IIHH, ICSI shortens the hormonal treatment and enhances the chances of pregnancy.

Seminal elastase-inhibitor complex, a marker of genital tract inflammation, and negative IVF outcome measures: role for a silent inflammation?

There is no consensus on whether and how male genital tract inflammation affects sperm fertilizing potential. The aims of this prospective study were to evaluate the elastase-inhibitor complex in seminal plasma (s-EI) level, a marker of male genital tract inflammation, in men undergoing in vitro fertilization (IVF) having no clinical signs of inflammation, and its association with semen characteristics, and the predictive role of s-EI for the outcome of IVF in terms of fertilization, embryo development to the blastocyst stage and pregnancy. The study involved 104 male partners of infertile couples with normal spermiogram undergoing IVF. On the day of oocyte retrieval, spermiogram and s-EI assessment using homogeneous immunoassay were performed. The outcome of IVF according to the s-EI level was assessed in 85 cycles with two or more oocytes. In 67 cycles embryos were cultured to the blastocyst stage. Spearman's and Pearson's correlation tests, chi-square test, and multiple linear and logistic regression analyses were used for statistical evaluation. Increased s-EI level (> or =250 microg/L) was detected in semen of 32% of men, and in 24% of men without leucocytospermia. We found an association between the s-EI level and semen leucocytes (r = 0.49, p = 0.004) but not with classical sperm characteristics. No correlation between the s-EI level and fertilization was observed. Increased s-EI levels were associated with a poorer blastocyst development rate (p = 0.03) and a higher number of arrested embryos (p = 0.04). Extended embryo culture to the blastocyst stage shows a negative effect of clinically silent male genital tract inflammation on embryo developmental potential.

Recombinant FSH vs. urinary FSH for ovarian stimulation in in vitro fertilization.

OBJECTIVE: To compare ovarian stimulation with recombinant FSH (rFSH) vs. urinary FSH (uFSH) in terms of hormonal events within ovarian follicles and the outcome of in vitro fertilization. STUDY DESIGN: A prospective randomized comparative study of rFSH (n = 70) vs. uFSH (n = 61) ovarian stimulation. Hormone determinations were serum estradiol (E2) on the day of human chorionic gonadotropin (hCG) administration, and E2, androstenedione (A) and testosterone (T) at the time of follicular aspiration in the follicular fluid and serum. RESULTS: The total dose of gonadotropins required and the length of ovarian stimulation were the same in the 2 groups. In follicular fluid the E2 and the A levels were significantly higher in the rFSH group (3,065 +/- 1,646 vs. 2,368 +/- 1,240 nmol/L, P = .004, and 103.7 +/- 51.6 vs. 89.0 +/- 42.3 nmol/L, P = .042, respectively), whereas A:E2 and T:E2 ratios were significantly lower (39.6 +/- 22.5 vs. 52.3 +/- 59.6, P = .042, and 9.1 +/- 4.7 vs. 17.6 +/- 26.9, P = .006, respectively). Serum hormonal levels, number of oocytes retrieved and pregnancy rates did not differ significantly between the groups. CONCLUSION: rFSH provides results similar to those of uFSH. rFSH enhances steroidogenesis and provokes different androgen/estrogen ratios than does uFSH without influencing the outcome of in vitro fertilization.

Seminal reactive oxygen species as predictors of fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization and intracytoplasmic sperm injection.

High seminal reactive oxygen species (ROS) are related to poor semen quality and impaired fertilization. We aimed at finding whether there is an association between ROS and fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In prepared semen of 147 male partners of infertile couples, ROS were assessed with luminol chemiluminescence. Spermiogram was assessed in native semen. ROS were negatively correlated with standard sperm characteristics and testicular volume, and positively with abnormal sperm head morphology. Fertilization rate and embryo morphology on day 2 and on day 4 were assessed in 41 IVF and 106 ICSI cycles. The influence of maternal (female age and number of oocytes) and paternal (sperm motility, morphology and ROS) factors on fertilization and embryo quality were assessed by means of regression analyses. After IVF, fertilization and pregnancy rates were negatively associated with ROS level (p = 0.031 and 0.041, respectively). In case of higher ROS, significantly fewer ICSI-derived embryos (p = 0.036) reached the morula-blastocyst stage on day 4. High seminal ROS levels are associated with impaired sperm fertilizing ability and lower pregnancy rates after IVF. In ICSI, a negative association of ROS with embryo development to the blastocyst stage has been observed.